超声微波协同提取药桑叶生物碱及其保肝活性研究

本研究以药桑叶为原料,采用单因素结合响应面法优化了药桑叶生物碱的超声微波协同提取工艺,并采用四氯化碳致小鼠急性肝损伤模型评价了药桑叶提取物(MNLE)的保肝活性。结果表明药桑叶生物碱的最佳提取工艺为:料液比1∶54 g/mL,乙醇浓度79%,微波功率300~0 W(0~7 min),超声功率493 W,提取时间29 min,提取温度70℃,在此条件下生物碱提取率可达11.325 mg/g。药效学试验结果显示,与模型组相比,MNLE能明显降低血清谷丙转氨酶(ALT)、谷草转氨酶(AST)水平以及肝组织丙二醛(MDA)和一氧化氮(NO)水平(P<0.05或P<0.01),升高肝组织超氧化物歧化酶(SOD)和谷胱甘肽(GSH)水平(P<0.05或P<0.01),肝组织病理学变化也得到了明显改善。结果表明MNLE具有较好的保肝作用,其作用机制与其抗氧化活性相关。

Development of a Total Flavonoids Extract of Artemisia Rupestris L.via Nanotechnology and Its Antiviral Effect In Vitro

Objective:We aimed to develop a novel Artemisia rupestris L.flavonoid nano-encapsulation(AFN)preparation and evaulate its anti-hepatitis B virus(HBV)activityin vitro.Methods:First,the AFN was prepared using polylactic-co-glycolic acid(PLGA).Then,after verification of the AFN,in vitro anti-virus assays were conducted by:(1)assessing the inhibitory effect of AFN on the secretion of hepatitis B surface antigens(HBsAg),hepatitis B e-antigens(HBeAg),and the replication of HBV DNAin HepG2.2.15 cells;(2)analyzing the influence of AFN on the activation rate of NF-κB positive cells;and(3)evaluating the effect of AFN on the function of glutathione peroxidase(GSH-PX)enzymes located onthe HepG2.2.15 cell membrane.Results:Compared to the original total flavonoids extract of Artemisia rupestris L.(withoutnano-encapsulation),AFN preparation under the maximum nontoxic concentration effectively inhibited the secretion of HBsAg,HBeAg,and HBV DNA from HepG2.2.15 cells.At the same time,AFN preparation promoted not only the activation rate of NF-κB positive cells,but also antiviral GSH-PX enzyme function.In conclusion,nano-encapsulation of the flavonoids extract of Artemisia rupestris L.showed an enhanced anti-HBV effect in vitro compared to the original total flavonoids extract(without nanoencapsulation);therefore,nano-encapsulation has great potential for the development of a novel antiviral herbal medicine preparation with improved efficacy.

秋菊丸对大鼠急性痛风性关节炎抗炎的实验研究

目的:观察秋菊丸对急性痛风性关节炎模型大鼠抗炎作用的影响。方法:SD雄性健康大鼠随机分为正常对照组、模型对照组、吲哚美辛8 mg/kg组及秋菊丸0.56、1.12、2.24 g/kg组,每组10只,各组灌胃相应药物或生理盐水连续12 d后,以Coderre法建立急性痛风性关节炎模型,造模后再灌胃2 d。分别在造模前、造模后6 h、12 h、24 h、36 h、48 h测量踝关节容积;于末次给药2 h后取大鼠踝关节滑膜组织;HE染色检查踝关节内滑膜的病理学变化及病理评分;ELISA法检测大鼠滑膜及关节腔冲洗液中肿瘤坏死因子-α(TNF-α)、白介素-1β(IL-1β)、IL-6的含量。结果:与正常对照组相比,各时相模型对照组关节肿胀度显著增加,大鼠滑膜组织中TNF-α、IL-1β、IL-6含量显著升高(P<0.01),镜下见模型对照组关节滑膜有大量炎症细胞浸润、异常血管及成纤维细胞增生,病理评分显著升高(P<0.01);与模型对照组比较,秋菊丸各剂量组在造模后6 h、12h、24 h、36 h、48 h关节肿胀度显著降低,TNF-α、IL-1β、IL-6含量显著降低,踝关节病理评分显著降低(P<0.01)。结论:秋菊丸可治疗大鼠急性痛风性关节炎,有显著抗炎作用,可能是通过抑制关节中TNF-α、IL-1β、IL-6炎性因子的释放,改善局部组织炎症反应。

秋菊丸化学成分鉴定及靶向GLUT9-OAT3尿酸转运体介导慢性高尿酸血症大鼠尿酸代谢和肾保护的机制研究

目的初步明确秋菊丸化学成分,研究秋菊丸对慢性高尿酸血症(HUA)模型大鼠尿酸(UA)升高及肾脏损伤的治疗作用及机制。方法借助超高效液相色谱串联四极杆静电场轨道阱质谱(UPLC-Q-Exactive Orbitrap MS)技术,初步鉴定秋菊丸的化学成分。48只雄性SD大鼠随机分成对照组、模型组、苯溴马隆(5.45 mg·kg^(-1),阳性药)组和秋菊丸低、中、高剂量(1.31、2.62、5.24 g·kg^(-1))组,除对照组外,通过ig腺嘌呤和乙胺丁醇制备大鼠慢性HUA模型,造模同时每天ig给药1次,连续28 d。使用全自动生化仪测定大鼠血清UA、肌酐(CRE)、尿素氮(BUN)水平;HE及Masson染色观察肾脏组织病理学变化;实时荧光定量PCR(qRT-PCR)法检测肾脏组织中葡萄糖转运蛋白9(GLUT9)、有机阴离子转运体家族蛋白3(OAT3)、ATP结合盒亚家族G成员2(ABCG2)、转化生长因子-β(TGF-β1)、SMAD蛋白3(Smad3)、α-平滑肌肌动蛋白(α-SMA)的mRNA水平。结果秋菊丸中共鉴定出41个成分,包括黄酮类(9个)、有机酸类(5个)、生物碱类(3个)、苯丙素类(7个)、香豆素类(1个)、酚类(3个)、呋喃类(2个)、脂肪酰类(6个)、醌类(2个)、萜类(1个)、内酯类(1个)、脂肪酸类(1个)。给药28 d后,HE染色结果表明,与模型组比较,各给药组肾损伤的病理学改变明显恢复,减轻了肾小球萎缩、肾小管扩张、肾纤维化及肾间质炎性细胞浸润等病理改变,其中秋菊丸高剂量组及苯溴马隆组肾脏损伤最轻。与模型组比较,秋菊丸高剂量组的CRE、UA水平均显著降低(P<0.01),各剂量组BUN水平有降低趋势,但无显著性差异;高、中剂量组肾组织中肾小管上皮细胞中胶原蛋白的含量显著降低(P<0.05、0.01);高剂量组GLUT9 mRNA表达量显著减少(P<0.05、0.01),各剂量组OAT3、ABCG2的mRNA表达量均显著增加(P<0.05、0.01);中、高剂量α-SMA mRNA表达量显著减少(P<0.05、0.01),高剂量组TGF-β1 mRNA表达量显著减少(P<0.01),各给药组Smad3的mRNA表达量有减少趋势,但无显著性差异。结论初步明确秋菊丸化成成分,秋菊丸具有增强UA代谢、减轻HUA所致肾脏损伤及肾纤维化的作用,其机制可能与调控GLUT9-OAT3尿酸转运体相关。

Protection of Citrullus colocynthis Fruit Extracts on Carbon Tetrachloride-induced and Bacillus Calmette-Guerin plus Lipopolysaccharide-induced Hepatotoxicity in Mice

Objective To investigate the hepatoprotective activities of the extracts from Citrullus colocynthis(ECC),a native plant used as traditional Uigur Medicine on acute liver injury in mice.Methods The activities of ECC of petroleum ether(ECCPE),chloroform(ECCC),ethyl acetate(ECCEA),n-butyl alcohol(ECCBA),and water(ECCW) were evaluated in vivo using two experimental models,carbon tetrachloride(CCl4)- and bacillus calmette-guerin(BCG) plus lipopolysaccharide(LPS)-induced acute hepatotoxicity in mice.The contents of aspartate aminotransferase(AST) and alanine aminotransferase(ALT) in serum were determined and the liver histological examination was carried out,respectively.Results The pretreatment with ECC for 7 d obviously reduced the impact of CCl4toxicity on the serum markers of liver damage,ECCEA and ECCC with a significant difference of AST(P < 0.01,0.05,respectively) and ALT(P < 0.05,0.01,respectively).The protective activity was reconfirmed against BCG + LPS-induced injury and the serum enzymatic levels were obviously elevated,for ECCEA and ECCC with a significant difference of AST(P < 0.05,0.01,respectively) and ALT(P < 0.01,0.05,respectively).Conclusion That ECCEA and ECCC are the potent hepatoprotective extracts that could protect liver against the acute injury,and this ability might be attributed to their hepatoprotective potentials.